Characterization of Biofilms by Correlated Mass Spectrometric and Raman Imaging

通过相关质谱和拉曼成像表征生物膜

• 批准号: 9064080
• 负责人: Joshua Shrout
• 金额: $ 43.8万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9064080
• 关键词: Address; Affect; Amino Acids; Animal Model; B-Lymphocytes; Bacteria; Bacterial Infections; Behavior; Biochemical; Cells; Characteristics; Chemicals; Clinical; Coculture Techniques; Communities; Complex; Detection; Development; Diagnostic; Dimensions; Escherichia coli; Extracellular Matrix; Gene Expression; Genes; Goals; Growth; Haemophilus influenzae; Health; Image; In Situ; In Vitro; Individual; Infection; Learning; Lung; Maps; Mass Spectrum Analysis; Measurement; Methodology; Methods; Microbe; Microbial Biofilms; Modeling; Molecular Profiling; Mucins; Nature; Population; Production; Pseudomonas; Pseudomonas aeruginosa; Quinolones; Regulation; Research; Resolution; Sampling; Signal Transduction; Signaling Molecule; Spatial Distribution; Surface; System; Therapeutic; Time; Variant; Virulence; Virulent; Work; design; extracellular; gene product; homoserine lactone; imaging modality; improved; insight; microbial; microbial community; pathogen; pathogenic bacteria; programs; quorum sensing; research study; rhamnolipid; signal processing; signature molecule; spectroscopic imaging; trait

DESCRIPTION (provided by applicant): Most bacteria in natural and clinical settings grow as surface-attached biofilms, which are bacterial communities that have self-assembled into an encased matrix of extracellular polymeric substances (EPS). To form these bacterial biofilm communities and infect host cells, an intercellular signaling process described as quorum sensing (QS) is very important. For the opportunistic pathogen, Pseudomonas aeruginosa, QS regulates the expression of many genes important to biofilm initiation, EPS production, and virulence. While much has been learned about select factors that regulate biofilm formation in vitro and in animal models, the specifics by which multispecies groups of cells form biofilms are not yet clear. Correlated mass spectrometric and Raman imaging will be used to develop a multiplex analysis method for studying host-associated microbial communities. These methods allow the determination of individual bacterial species and their microbial products within a mixed bacterial community growing in situ on surfaces. A long-term goal is the design of detection and diagnostic strategies informed by an understanding of bacterial interactions and signature biomolecule production. Work toward this goal will begin by conducting experiments with the opportunistic pathogen Pseudomonas aeruginosa, specifically developing methods to distinguish P. aeruginosa from other bacterial species, distinguish separate P. aeruginosa strains from each other, and determine the spatial distribution of four types of P. aeruginosa molecules associated with virulence: acyl- homoserine lactones (AHL), quinolones, such as pseudomonas quinolone signal (PQS), rhamnolipids, and exopolymeric substances (EPS). Successfully implementing this research program will result in bacterial mapping methods with improved sensitivity and resolution, substantially enhancing our ability to identify specific species of bacteria in mixed samples, and to identify and map specific chemical products produced by these bacteria that are critical to host colonization, infection, and virulence.

描述（由申请人提供）：自然和临床环境中的大多数细菌以表面附着生物膜的形式生长，生物膜是自组装成细胞外聚合物（EPS）包裹基质的细菌群落。为了形成这些细菌生物膜群落并感染宿主细胞，被称为群体感应（QS）的细胞间信号传导过程非常重要。对于条件致病菌铜绿假单胞菌，QS调节许多对生物膜起始、EPS产生和毒力重要的基因的表达。虽然在体外和动物模型中对调节生物膜形成的选择因素已经了解了很多，但多物种细胞群形成生物膜的具体情况尚不清楚。 相关的质谱和拉曼成像将被用来开发一个多重分析方法，研究主机相关的微生物群落。这些方法允许确定在表面上原位生长的混合细菌群落中的单个细菌物种及其微生物产物。 长期目标是通过了解细菌相互作用和特征生物分子产生来设计检测和诊断策略。实现这一目标的工作将开始，首先用机会致病菌铜绿假单胞菌进行实验，特别是开发区分铜绿假单胞菌与其他细菌物种的方法，区分不同的铜绿假单胞菌菌株，并确定与毒力相关的四种类型的铜绿假单胞菌分子的空间分布：酰基高丝氨酸内酯（阿勒）、喹诺酮类，如假单胞菌喹诺酮信号（PQS）、鼠李糖脂和外聚合物（EPS）。成功实施这项研究计划将导致细菌作图方法具有更高的灵敏度和分辨率，大大提高我们在混合样品中识别特定细菌物种的能力，并识别和绘制这些细菌产生的对宿主定植，感染和毒力至关重要的特定化学产物。